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Different pharmacological anatomy in the paraventricular hypothalamic nucleus, supraoptic nucleus, and suprachiasmatic nucleus of rats: quantitative autoradiography on angiotensin II receptor binding sites.

Angiotensin II (AII) and vasopressin (VP) play important roles in cardiovascular function. Using 125I-[Sar1,Ile8]-angiotensin II (125I-SI-AII), a potent AII antagonist, AII receptor binding sites were autoradiographically localized in three VP-producing areas of the hypothalamus and compared in hypertensive and normotensive rats. Within three major VP-producing areas, AII receptor binding was highest in the paraventricular hypothalamic nucleus and lowest in the supraoptic nucleus, suggesting that a differential AII regulation of separate VP systems exists in the brainstem. No statistical difference in 125I-SI-AII receptor binding was found between WKY and SHR rats in each of the three major VP-producing nuclei studied. These results are consistent with a role of AII receptors in a subtle and complicated regulation of VP in cardiovascular function.

Angiotensin II↗

The effects of estrogen and progesterone on corticotropin-releasing hormone and arginine vasopressin messenger ribonucleic acid levels in the paraventricular nucleus and supraoptic nucleus of the rhesus monkey.

Ovarian steroids increase hypothalamic-pituitary-adrenal (HPA) axis activity and sensitize the hypothalamic-pituitary-ovarian (HPO) axis to stress-induced inhibition. The present study investigated the effect of ovarian steroids on CRH and arginine vasopressin (AVP) messenger RNA (mRNA) levels in the rhesus monkey hypothalamus, as both neuropeptides have been shown to stimulate the HPA axis and inhibit the HPO axis in this species. This was accomplished by measuring CRH and AVP mRNA in the paraventricular nucleus (PVN) and supraoptic nucleus (SON) by in situ hybridization histochemistry. Menstrual cycles were simulated in ovariectomized (OVX) rhesus monkeys by sequential addition and removal of SILASTIC brand (Dow Corning Corp.) tubing containing either 17beta-estradiol (E2) or progesterone (P4). On the morning of day 11 of the simulated follicular phase (E2 alone) or day 21 of the luteal phase (E2 + P4), animals were anesthetized, and the brains were perfused with paraformaldehyde via the carotid artery. Coronal sections (30 microm) were cut, and mRNA for CRH and AVP in the paraventricular nucleus (PVN) and supraoptic nucleus (SON) were semiquantified by in situ hybridization. CRH mRNA in the PVN of E2-replaced OVX animals (n = 7) was 2-fold greater than that in untreated OVX controls (n = 4), whereas CRH mRNA after E2 + P4 (n = 4) was no different from that in controls (optical density + SEM, 0.38 +/- 0.06, 0.13 +/- 0.08, and 0.14 +/- 0.09 for OVX + E2, OVX + E2 + P4, and OVX, respectively; P = 0.02). CRH in the SON was undetectable. In contrast to CRH, AVP mRNA in the PVN and the SON was similar in the three treatment groups. We conclude that E2 and E2 + P4 replacement to OVX monkeys exert different effects on CRH and AVP gene expression, as estrogen stimulation of CRH mRNA in the PVN was abrogated by progesterone, whereas no effect of ovarian steroids on AVP mRNA in either the PVN or SON was observed. We postulate that ovarian steroid regulation of CRH synthesis and release may in part explain the central nervous system mechanisms by which ovarian steroids affect the HPA and HPO axes during basal and stress conditions.

Animals↗

Opposite regulation of body temperature by cholinergic input to the paraventricular nucleus and supraoptic nucleus in rats.

Hypothalamic cholinergic system plays an important role in the regulation of body temperature and fluid balance. We have previously shown that cholinergic stimulation of the anterior hypothalamus and preoptic area was accompanied by a fall in body temperature, increased water intake, and increased Fos protein in the paraventricular nucleus (PVN) and supraoptic nucleus (SON). In the present study, to estimate the role played by cholinergic input to the PVN and SON in thermoregulation and water intake, we used microdialysis for cholinergic stimulation with neostigmine and analysis of the nucleus, and also investigated immunoreactivity for c-Fos protein in the brain. This stimulation increased extracellular concentration of acetylcholine in these nuclei. Stimulation of the PVN decreased body temperature and increased water intake. On the other hand, stimulation of the SON increased body temperature. Both in PVN-stimulated and SON-stimulated rats, c-Fos-like immunoreactivity (Fos-IR) was evident in the PVN, SON and certain regions including locus coeruleus (LC), area postrema and nucleus of the solitary tract (NTS). Addition of atropine to the dialysis medium attenuated the increase of Fos-IR and suppressed the cholinergic stimulation-induced responses in body temperature and water intake. These results suggest that cholinergic muscarinic mechanisms in PVN and SON play an opposite function in the regulation of body temperature. The same neuronal pathway including LC and NTS may participate in an advance both in hypothermia and in hyperthermia.

Acetylcholine↗

GABAergic projection from the arcuate nucleus to the supraoptic nucleus in the rat.

Electrical stimulation of the neurones in the hypothalamic arcuate nucleus results in a transient inhibition followed by a marked post-stimulus excitation of magnocellular neurones of the supraoptic nucleus. Microdialysis administration of the gamma-aminobutyric acid agonist (GABA(A)), muscimol, directly into the supraoptic nucleus inhibited both oxytocin and vasopressin neurones and these actions were fully reversed by the GABA(A) antagonist bicuculline. In addition, bicuculline administration blocked the inhibition induced by arcuate stimulation, but had no effect on the post-stimulus excitation. Thus, part of the inhibitory pathway arising from or passing through the arcuate nucleus to the supraoptic nucleus is mediated by the neurotransmitter GABA. However, the post-inhibitory excitation induced by arcuate stimulation is not a rebound response, but appears to involve an independent excitatory pathway.

Animals↗

Magnocellular tuberomammillary nucleus input to the supraoptic nucleus in the rat: anatomical and in vitro electrophysiological investigations.

Anatomical and electrophysiological methods were used to investigate the existence and role of inputs from the magnocellular tuberomammillary nucleus to the supraoptic nucleus. After injecting either Fluoro-Gold or rhodamine-labeled latex microspheres into the supraoptic nucleus, consistent patterns of retrogradely labeled neurons within the tuberomammillary nucleus were observed. The results indicate that both subdivisions of the supraoptic nucleus, the tuberal and the anterior, receive input from the tuberomammillary nucleus. Injections into the tuberal supraoptic nucleus tended to label more cells in the contralateral tuberomammillary nucleus, while injections into the anterior supraoptic nucleus may label more cells on the ipsilateral side. The in vitro intracellular electrophysiological results support the anatomical findings and extend them in several ways. Some tuberomammillary neurons were found to project to the supraoptic nuclei on both sides of the brain. Intracellular Lucifer Yellow injections into tuberomammillary cells after electrophysiological recording revealed labeled axons that were traceable into the supraoptic nucleus, where apparent varicosities (possible en passant terminals) were seen. Magnocellular tuberomammillary nucleus neurons had characteristic passive and active membrane properties and morphology, similar to histaminergic neurons in this area studied by other workers. Finally, in two of the 21 cases, Lucifer Yellow injection into one neuron revealed dye-coupled pairs of tuberomammillary neurons. Previous work by others has shown that histamine excited cells in the tuberal subdivision of the supraoptic nucleus, stimulating vasopressin release, and that the tuberomammillary nucleus provides histaminergic input to the anterior portion of the supraoptic. The present findings show that the tuberomammillary nucleus supplies input to both subdivisions of the supraoptic nucleus and that this input is provided bilaterally. Taken together with previous work, these data suggest that the tuberomammillary nucleus provides histaminergic input to the supraoptic nucleus and may be involved specifically with vasopressin release.

Action Potentials↗

Electrophysiological evidence for a projection from the arcuate nucleus to the supraoptic nucleus.

The electrical activity of single neurones in the hypothalamic arcuate and supraoptic nuclei was recorded in urethane-anaesthetized rats. Stimulus pulses applied to the supraoptic nucleus antidromically activated 3 out of 41 cells recorded in the ipsilateral arcuate nucleus, confirming that there is a projection from the arcuate nucleus to the region of the supraoptic nucleus. Stimulation of the arcuate nucleus inhibited 17 out of 19 continuously firing (putative oxytocin) supraoptic neurones. Inhibition was followed by a marked post-stimulus excitation in 12 cells. The responses were not abolished by i.v. injection of the opioid antagonist naloxone. Thus at least part of the input to the magnocellular oxytocin system that arises from or passes through the arcuate nucleus, is not mediated by opioid peptides.

Animals↗

Beta-endorphin cells in the arcuate nucleus: projections to the supraoptic nucleus and changes in expression during pregnancy and parturition.

Supraoptic nucleus oxytocin neurone activity and secretion are inhibited in late pregnancy and parturition by endogenous opioids. Here, we investigated alterations in the projections and gene expression of beta-endorphin/pro-opiomelanocortin neurones in the arcuate nucleus in the pregnant rat. All regions of the arcuate nucleus were found to contain cells immunoreactive for beta-endorphin fluorescent microbeads retrogradely transported from the supraoptic nucleus, and double-labelled neurones (beta-endorphin plus microbeads), showing that beta-endorphin neurones throughout the arcuate nucleus project to the supraoptic nucleus. There was an increase in the number of beta-endorphin-immunoreactive cells in the arcuate nucleus and an increase in the density of beta-endorphin fibres within the supraoptic nucleus and peri-supraoptic region in late pregnancy and parturition, suggesting enhanced expression of beta-endorphin and increased beta-endorphin innervation of the supraoptic nucleus. Pro-opiomelanocortin mRNA expression in the arcuate nucleus increased in late compared to early pregnancy: the number of positive neurones significantly increased in the caudal region. Fos expression (an indicator of neuronal activation) in the arcuate nucleus was colocalized in beta-endorphin neurones in both proestrus and parturient rats, but the number of positive cells did not increase during parturition, suggesting lack of activation of beta-endorphin neurones at birth. Thus, beta-endorphin cells in the arcuate nucleus project to the supraoptic nucleus and increased innervation during pregnancy may explain the enhanced endogenous opioid inhibition of oxytocin neurones.

Animals↗

Sex-specific effects of met-enkephalin treatment on vasopressin immunoreactivity in the rat supraoptic nucleus.

The supraoptic nucleus of male and female rats treated with met-enkephalin or naloxone and met-enkephalin was examined with light microscopical immunocytochemistry for Arginine-vasopressin. Both genders exhibited the same distribution of immunostained magnocellular neurons. Met-enkephalin treatment caused an increase in number of immunostained vasopressin neurons. This effect was more pronounced in females than in males. Naloxone treatment diminished immunoreactive cytoplasmic vasopressin in males more effectively than in females. In enkephalin-treated animals numerous vasopressin immunoreactive varicosities appeared within the supraoptic nucleus, but were mostly absent in naloxone-treated animals and in controls. Our results indicate that met-enkephalin treatment either stimulates vasopressin synthesis or inhibits secretion. It is likely that steroid hormones mediate the action of enkephalin on vasopressin secretion in a specific manner.

Animals↗

Differences in function and structure of the capillary endothelium in the supraoptic nucleus and pituitary neural lobe of rats. Evidence for the supraoptic nucleus as an osmometer.

The physiology and structure of capillary endothelial cells in the hypothalamic ventromedial and supraoptic nuclei and pituitary neural lobe were evaluated with quantitative methods and compared. The capillary endothelial cells in the ventromedial nucleus were used as an index of blood-brain barrier endothelium in cerebral gray matter; this endothelium has relatively low surface area and low permeability to tracer solutes. The permeability X surface area product of endothelial cells for a neutral amino acid, 14C-alpha-aminoisobutyric acid (AIB), in the ventromedial nucleus was similar to the value for supraoptic nucleus and was several hundred times smaller than in the neural lobe. The supraoptic nuclei and neural lobe had exceptionally large capillary surface areas, but dissimilar rates of blood flow and transendothelial influx of AIB. Differences in permeability of the endothelial cells between these two structures correlated closely with their marked dissimilarities in morphology. The neural lobe endothelium had numerous fenestrations (five per capillary cross-section) and vesicular profiles (twice as many as supraoptic nucleus), two features commonly associated with high capillary permeability. The capillary endothelium of the supraoptic nucleus was that of a typical blood-brain barrier structure having intercellular junctions that appeared tight, no fenestrations, and few cytoplasmic pits and vesicles. The unusually large capillary surface area of the supraoptic nucleus and low rate of solute flux across its endothelial cells make this nucleus a unique structure in which rapid changes in tissue volume may occur in response to small perturbations in plasma osmolality. The findings implicate the supraoptic nucleus as an osmotically sensitive detector or 'osmometer' in neuroendocrine regulation of body fluid homeostasis.

Animals↗

Involvement of the noradrenergic afferents from the nucleus tractus solitarii to the supraoptic nucleus in oxytocin release after peripheral cholecystokinin octapeptide in the rat.

Activation of abdominal vagal afferents by peripheral injection of cholecystokinin octapeptide induces oxytocin release into the circulation. To test the hypothesis that cholecystokinin increases oxytocin release via activation of noradrenergic afferents from the brainstem, we injected rats with 5-amino-2,4-dihydroxy-alpha-methylphenylethylamine, a selective neurotoxin to noradrenergic fibres, into a lateral cerebral ventricle. The neurotoxin treatment reduced the noradrenaline content in the hypothalamus by 75% and reduced the oxytocin secretion in response to cholecystokinin by over 90%. In separate experiments, the neurotoxin was injected unilaterally in the vicinity of the supraoptic nucleus to test whether direct noradrenergic afferents to the supraoptic nucleus are involved in the response to cholecystokinin. The injection reduced the immunoreactivity for dopamine beta-hydroxylase in the supraoptic nucleus and significantly decreased the number of the supraoptic neurons expressing Fos-like protein after cholecystokinin but not after hypertonic saline. In further experiments, rhodamine-conjugated latex microspheres were injected into the supraoptic nucleus to retrogradely label afferent neurons, and the brains were processed with double-immunohistochemistry for tyrosine hydroxylase and Fos-like protein. In the C2/A2 but not the C1/A1 region of the brainstem, cholecystokinin increased the expression of Fos-like protein in the population of retrogradely-labelled catecholaminergic cells. In the C2/A2 region, the majority of retrogradely labelled cells expressing Fos-like protein after cholecystokinin were catecholaminergic. We conclude that noradrenergic afferents from the A2 but not from the A1 region of the brainstem to the hypothalamus mediate, at least in part, oxytocin release following cholecystokinin.

Afferent Pathways↗

Direct catecholaminergic projection from nucleus tractus solitarii to supraoptic nucleus.

To determine whether the supraoptic nucleus (SON) receives a direct projection from catecholamine cells of the nucleus tractus solitarii (NTS), retrograde transport of rhodamine-tagged latex microspheres was combined with a procedure for the fluorescence histochemical visualization of catecholamines. SON tracer injections, made via transpharyngeal approach, retrogradely labelled cells at all levels of NTS, although the majority were located caudal to obex with an ipsilateral predominance. Approximately half of these cells were also identified as catecholaminergic; the relatively caudal level in the dorsomedial medulla of most of these cells suggests that they probably correspond to the A2 catecholamine cell group.

Animals↗

A rat brain slice preserving synaptic connections between neurons of the suprachiasmatic nucleus, organum vasculosum lamina terminalis and supraoptic nucleus.

The organum vasculosum lamina terminalis (OVLT), the suprachiasmatic nucleus (SCN) and the supraoptic nucleus (SON) are three hypothalamic structures involved in the osmotic and circadian control of neurohypophysial secretion. Recent experiments have suggested that interactions between osmotic and circadian factors may be important for homeostasis. The existence of an in vitro slice preparation retaining these nuclei and their interconnections would therefore be useful for the analysis of synaptic interactions. In the rat, the OVLT, SCN and SON are found at increasingly ventral and lateral positions along the rostro-caudal axis, such that conventional 400 microm slices taken in the pure coronal or horizontal planes do not retain all three nuclei. Here we show that horizontal slices cut at angles of 38-42 degrees relative to the dorsal surface of the cortex retain large fractions of the three nuclei. Intracellular recordings revealed membrane properties consistent with those previously published for OVLT, SCN and SON neurons. Moreover, antidromic and synaptic responses evoked by electrical stimulation revealed that extensive axonal projections are retained between these nuclei. Finally, chemical and osmotic stimulation of the OVLT exerted powerful influences on the rate of spontaneous synaptic events in SON neurons. We therefore conclude that angled horizontal hypothalamic slices represent a useful preparation for the analysis of physiological interactions between the OVLT, SCN and SON.

Animals↗

Electrophysiologic evidence for neural connections between the paraventricular nucleus and neurons of the supraoptic nucleus in the rat.

These experiments were part of a study to examine the extrahypothalamic projections of the paraventricular nucleus in the rat. Thirty-nine cells, histologically identified as situated within the supraoptic nucleus, were recorded. Nine of those cells were classified as secreting vasopressin by their phasic patterns of electrical discharge. Six of the phasically firing cells were excited after electrical stimulation of the paraventricular nucleus and the other three were inhibited. Eight cells were antidromically identified as projecting to the paraventricular nucleus; most of these were not spontaneously active. Of the remaining 22 cells, the majority were spontaneously active, showing continuous patterns of firing, and most were inhibited after stimulation of the paraventricular nucleus. The data supply evidence for the presence of functional neural connections between the two major nuclei of the tuberohypophyseal neurosecretory system.

Animals↗

Parabrachial nucleus projection towards the hypothalamic supraoptic nucleus: electrophysiological and anatomical observations in the rat.

It has been proposed that the pontine parabrachial nucleus (PBN) participates in the regulation of body fluid balance and the release of vasopressin from the neurohypophysis, although the pathways mediating the latter response are uncertain. This study in the rat, utilizing anatomical and electrophysiological methods, describes a projection from the lateral PBN towards the hypothalamic supraoptic nucleus (SON). Rats received iontophoretic injections of the anterograde tracer Phaseolus vulgaris leucoagglutinin (PHA-L, 2% solution). After 14-17 days, rats were sacrificed and their brains prepared for immunofluorescent visualization of projections to the SON region. PHA-L-labelled terminals were found primarily in perinuclear regions immediately dorsal to the SON. In contrast, injections within the medial PBN and the nearby Kölliker-Fuse nucleus did not reveal labelling in or around the SON. Extracellular recordings from 86 of 118 antidromically identified neurons in anaesthetized rats revealed a set of complex synaptic responses after stimulation in the PBN. Excitatory responses (in 82 of 86 cells) of short (less than 100 msec, 39/82 cells) and long (greater than 100 msec, 43/82) duration were observed in both vasopressin- and oxytocin-secreting cells of the SON, while 4/86 cells displayed a depressant response to PBN stimulation. In the adjacent perinuclear zone, 22/39 nonneurosecretory cells responded with an increase in their excitability consequent to an identical stimulus. These data suggest a predominantly facilitatory influence of lateral PBN neurons on SON neurosecretory cells in the rat, and that the PBN-SON projection is an indirect one that utilizes an interneuronal network located in the perinuclear zone adjacent to the SON.

Animals↗

Nicotine facilitates noradrenaline release in the rat supraoptic nucleus.

Nicotine injected in the supraoptic nucleus facilitates vasopressin release from the neurohypophysis. Nicotinic acetylcholine receptors have been found not only on vasopressin-producing cell bodies but also on presynaptic nerve terminals in the nucleus. Vasopressin cells receive excitatory synaptic inputs from noradrenergic neurones. To test whether nicotine facilitates noradrenaline release in the supraoptic nucleus, we perfused the supraoptic nucleus with nicotine through a microdialysis probe. Nicotine increased the extracellular noradrenaline concentrations in the nucleus. A noradrenaline uptake inhibitor, desipramine, increased the extracellular noradrenaline concentrations in the nucleus and did not block the noradrenaline increase after nicotine. The results suggest that nicotine acts within the supraoptic nucleus to facilitate noradrenaline release pre-synaptically. This presynaptic action may contribute, in part, to vasopressin release after nicotine.

Anesthesia↗

Peripartum interneuronal coupling in the supraoptic nucleus.

In the supraoptic nucleus (SON), the incidence of conducting gap junctions (gjs), as indicated by dye coupling, is low in cycling females, but dramatically elevated in nursing mothers. Functionally, this is consistent with the well-established presence of synchronous milk ejection bursts among oxytocin neurons only in the lactating rat. In situ hybridization data, however, revealed elevated gj mRNA expression on the last day of pregnancy, a time when burst firing by putative oxytocin neurons is absent. Using Lucifer Yellow dye coupling, we determined the incidence of high conductance gjs in SONs of proestrous, immediately prepartum, postpartum non-lactating, lactating day 1, and lactating day 9-10 rats. Results indicate that coupling incidence is high only at times when milk ejection bursts are known to occur, and that the elevated gj mRNA expression seen on the last day of pregnancy does not indicate conducting gjs. It is suggested that gj conductance states, but not gj expression, are modulated by plasma estradiol titers.

Action Potentials↗

Sex- and age-related P75 neurotrophin receptor expression in the human supraoptic nucleus.

The human supraoptic nucleus (SON) is the main production site of plasma vasopressin. Previously, using the Golgi apparatus and cell size as measures for neuronal metabolic activity, an activation of vasopressinergic neurons was found during ageing in the human SON in women but not in men. We hypothesized that the low-affinity neurotrophin receptor p75 (p75(NTR)) might be involved in the mechanism of activation of vasopressin neurons in postmenopausal women, since this receptor was found to be expressed in the SON neurons of aged individuals, and because p75(NTR) expression was shown to be suppressed by estrogens. Therefore, we investigated whether p75(NTR) immunoreactivity in the SON neurons was age- and sex-dependent. For this purpose, we studied paraffin sections of the SON in 32 postmortem brains of control patients ranging in age from 29 to 94 years with an anti-p75(NTR) antibody and determined the area of p75(NTR) immunoreactivity per neuron using an image analysis system. To study whether the p75(NTR) might also participate in the activation of SON neurons, we related Golgi apparatus size to the area of p75(NTR) immunoreactivity per cell in the same patients. We found that the area of p75(NTR) immunoreactivity per cell correlated indeed significantly with age and with Golgi apparatus size only in women but not in men. Therefore, our results suggest that p75(NTR) is involved in postmenopausal activation of vasopressinergic neurons in the human SON.

Adult↗